植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03对TNBS诱导小鼠结肠炎的缓解作用

目的 为阐明益生菌抗氧化与结肠炎的关系,对植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03缓解三硝基苯磺酸（trinitro-benzene-sulfonic acid,TNBS）诱导的小鼠结肠炎进行探究。方法 通过对BALB/c小鼠肛门注射TNBS,构建小鼠结肠炎模型;分别采用植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03的单菌悬液（109 CFU/mL）及1∶1混合菌悬液（109 CFU/mL）进行8 d灌胃治疗。结果 治疗组小鼠结肠组织炎性细胞浸润症状获得缓解,血清中谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-PX）（t1=3.247,P1<0.05;t2=3.397,P2<0.05）、过氧化氢酶（catalase,CAT）（t1=5.289,P1<0.001;t2=3.563,P2<0.05）和总超氧化物歧化酶（total superoxide dismutase,T-SOD）（t1=3.317,P1<0.05;t2=3.551,P2<0.05）活性均有显著恢复。结论 植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03可通过增强机体抗氧化酶活性,起到缓解TNBS诱导的小鼠结肠炎的作用。     

两歧双歧杆菌86321生长特性的研究

目的研究两歧双歧杆菌86321的生长特性,为该菌生理功能研究和高效发酵剂的研制提供理论依据。方法通过生长曲线、产酸量、最适厌氧方式、最适pH、最适培养温度及最适接种量等一系列实验,对两歧双歧杆菌86321进行生长特性的研究。结果两歧双歧杆菌86321在BL培养基中培养时间可缩短至16 h,最高活菌数的lg值达到9.5;其最适厌氧方式为自然厌氧法或密封法,装液量视实际情况而定;在pH7.08.0生长良好,最适初始pH为8.0;在3742℃生长良好,最适温度为37℃;综合总菌量和生产成本,确定最适接种量为7%（v/v）。结论用BL培养基可以大大提高两歧双歧杆菌86321的产量。细菌产量的高低和发酵速度的快慢与菌种活力、厌氧方式、培养温度及pH等因素密切相关。     

乳杆菌和双歧杆菌与人类健康的关系

乳杆菌和双歧杆菌与人类健康的关系原解放军３０９医院北京１０００４５何道生１前言为了健康的目的人类利用乳杆菌的历史可以追朔到１９０８年。细菌学的创始人梅奇尼可夫提出,“为了延长人类的生命,应该饮用乳杆菌发酵过的牛奶。”他说,肠道微生物能产生“毒素”,使...     

双歧杆菌和乳杆菌在诱发抗肿瘤免疫中的作用

双歧杆菌和乳杆菌给封闭群昆明小鼠腹腔注射,在体内激活后,胸腺细胞和脾细胞对ＣｏｎＡ刺激的增殖反应,脾贴附性细胞对ＹＡＣ－１,Ｌ９２９的细胞毒作用,以及脾贴附性细胞产生对上述二株瘤细胞的肿瘤坏死因子（ＴＮＦ）的活性都比对照动物明显增强。结果提示短双歧杆菌和嗜酸性乳杆菌给小鼠腹腔注射后,通过激活脾脏淋巴细胞和贴附性细胞（巨噬细胞）所介导的免疫功能而明显地增强宿主的抗肿瘤活性。     

双歧杆菌与乳杆菌原生质体的融合及筛选

目的：通过双歧杆菌与乳杆菌的原生质体融合构建耐氧双岐杆菌,以解决双岐杆菌制剂开发中始终存在的“活菌数低”和存活时间短等问题。方法：采用不同浓度的Mutanolysin分别制备长岐杆菌和保加利亚乳杆菌的原生质体,在电场作用下诱导长双岐杆菌原生质体和经56℃热灭活30min的保加利亚乳杆菌原生质体融合,并在双层再生培养基上筛选融合菌株。结果：成功地构建出几种较为稳定的兼具长双岐杆菌和保加利亚乳杆菌生物学特性的融合菌株;其中融合株F2具有良好的耐氧性;且能在有氧、CO2条件下良好生长。结论：通过原生质体融合技术能改良双歧杆菌的严格厌氧特性,有利于对双岐杆菌的开发和应用。     

双歧杆菌、乳杆菌制剂在食品工业中的研究现状与展望

正常人体肠道中存在着大量微生物 ,这些菌群的平衡对人体健康发挥着重要的作用。肠道内正常菌群作为一个整体存在 ,相互依赖又相互制约。机体肠道内菌群失调则出现一系列的生理功能紊乱 ,临床表现为腹泻和感染等。而近年来 ,益生菌 (剂 ) (probiotics)作为一类对人体有益的细菌的总称 ,它对腹泻和感染等疾病的治疗和预防有特殊的功效 ,已成为微生物学的研究热点 ,并在保健食工业上得到广泛应用。双歧杆菌、乳杆菌是目前开发应用最热门的两种益生菌。双歧杆菌、乳杆菌是机体有益的与人类健康又密不可分的重要生理菌群 ,二者对人健康和营养有…     

螺旋藻在体外对双歧杆菌及乳杆菌增殖的促进作用

通过用改良的ＴＰＹ和ＧＡＭ培养基作为螺旋藻的基质,在试管内证实了螺旋藻对双歧杆菌、乳杆菌生长的促进作用。结果显示,钝顶螺旋藻在实验浓度下,对双歧杆菌和乳杆菌的增殖作用与对照组相比有非常显著的差异,这一结果也说明了钝顶螺旋藻在调整肠道菌群,改善肠道内环境方面的作用。     

乳杆菌及双歧杆菌缓解便秘的机制

探讨可改善患者便秘症状的植物乳植杆菌LP45、嗜酸乳杆菌La28和动物双歧杆菌乳亚种BAL531的作用机制。

采用HT-29细胞与肠道菌群体外批量发酵系统结合的方法,检测益生菌增殖肠上皮细胞、提升肠道紧密连接性与黏膜屏障能力以及增强五羟色胺转运体（SERT）、水通道蛋白-3（AQP-3）表达的能力。

菌株LP45和BAL531均具有较好的黏附能力并能够增殖肠上皮细胞,菌株La28无增殖作用;菌株LP45、La28和BAL531能显著提升紧密连接蛋白ZO-1和Occludin的mRNA相对表达水平,最高达2.73倍（t = 13.099,P＜0.001）,从而增强肠道屏障。菌株LP45使黏蛋白（MUC2）mRNA相对表达量显著提升1.89倍（t = 10.285,P = 0.001）,菌株La28与BAL531无显著影响;菌株LP45、La28和BAL531使SERT mRNA相对表达量分别提升2.13倍、1.45倍和4.00倍,其中菌株BAL531效果最显著（t = 21.308,P＜0.001）;另外,菌株LP45可使AQP-3 mRNA相对表达量提升2.12倍（t = 10.625,P＜0.001）,菌株BAL531使AQP-3 mRNA相对表达量下调0.79倍（t = ‒2.611,P = 0.059）,菌株La28无显著影响（t = ‒0.951,P = 0.395）。

菌株LP45、La28和BAL531对便秘症状的缓解作用可能是通过特异性增加肠道润滑、促进肠道蠕动、减缓肠道水液吸收等不同机制实现的。

     

稀土元素对两歧双歧杆菌生长的影响

目的：报道不同浓度稀土元素铈和镧（1－1000ug/ml)对两歧双歧杆菌生长的影响。方法：试管培养与连续厌氧培养法,结果：当两种稀土元素浓度大于300ug/ml时,双歧杆菌的生长明显受到抑制,当稀土元素浓度在100－200ug/ml时,对双歧杆菌生长有轻微抑制,低浓度的稀土元素对双歧杆菌无刺激生长作用,稀土元素（100ug/ml)对厌氧连续培养的双歧杆菌同样有明显的抑制作用,随着稀土元素被洗脱,细胞的生长才逐渐缓慢恢复,试管培养中加入一定浓度稀土元素（100ug/ml),连续取样检查,也可观察到稀土元素对双歧杆菌的抑制情况,结论：一定浓度稀土元素铈和镧对双歧杆菌生长有促进作用。     

双歧杆菌与乳杆菌对大鼠肠菌群失调的调整作用的初步研究

本试验初步研究了双歧杆菌ＤＭ９２２７菌株及乳杆菌ＤＭ８１２１菌株对大鼠肠菌群失调的调整作用。将含有红霉素及链霉素的抗生素溶液给２０只大鼠灌胃,灌胃量为每种抗生素２００ｍｇ／ｋｇ／日,连续３天。然后将动物分为双歧杆菌及乳杆菌菌液灌胃组及ＰＢＳ灌胃对照组。     

1株新分离的人两歧双歧杆菌耐药性研究

目的研究新分离的人两歧双歧杆菌对11种抗生素的耐药性。方法采用琼脂扩散纸片法测定人两歧双歧杆菌对11种抗生索的敏感性,通过在双歧杆菌培养基中添加不同浓度的抗生素来确定MIC值。结果该菌株对β-内酰胺类、大环内脂类和利福平非常敏感．对氨基糖苷类、黄胺类表现出较强抗性。结论该菌株对11种抗生索的药物敏感性与国内外其他文献所报道的结果一致。但该菌株带有一22kb大小的天然质粒。需进一步研究质粒与其耐药性的关系。     

Effects of Bifidobacterium bifidum on adaptive immune senescence in aging mice

Bifidobacteria are a natural component of the bacterial flora of the human body and have a symbiotic bacteria-host relationship with human beings. Aging is associated with reduced numbers of beneficial colonic Bifidobacteria and impaired immunity. The possible anti-senescence effects of Bifidobacteria are presently unknown. The aims of the present study were to investigate possible anti-senescence effects of B. bifidum on naturally senescent mice and to explore their mechanisms. After treatment with B. bifidum, mice were killed and samples collected. Cytokine production in serum and lymphocyte culture supernatant, anti-oxidation activity and gene expression were measured. B. bifidum significantly increased cytokine IL-2 and IFN-γ levels but decreased proinflammatory cytokines IL-6 and TNF-α concentrations. Moreover, B. bifidum improved anti-oxidation activity and reduced lipid peroxidation in thymus and spleen. In addition, B. bifidum down-regulated p16 expression in thymus and spleen. Taken together, the results indicate, for the first time, that B. bifidum delays senescence by several mechanisms, including enhancement of anti-oxidation activity in thymus and spleen, alteration of gene expression and improvement in immune function.     

低pH处理对两歧双歧杆菌KLDS2.0603黏附能力的影响

【目的】确定低pH处理对两歧双歧杆菌KLDS2.0603黏附能力及其表面物理化学性质的影响。【方法】将两歧双歧杆菌KLDS2.0603菌体在不同低pH的PBS溶液中处理一定时间后,采用平板菌落计数法和直接镜检法,测定其经历不同pH的酸性环境后的黏附能力,及其表面疏水性和自动聚集能力。【结果】不同pH的PBS溶液处理后的双歧杆菌菌体,其黏附能力均不同程度下降,除pH 5.0的处理组外,其余处理组均显著低于空白组。此外,经不同pH的PBS溶液处理后,仅pH 3.0和3.5的两处理组,双歧杆菌表面疏水性显著提高。除pH 1.0、1.5和5.0的处理组外,其余处理组的自动聚集能力均显著下降。【结论】低pH的酸性环境会降低两歧双歧杆菌KLDS2.0603的黏附能力,并且双歧杆菌的自动聚集能力和表面疏水性也发生相应变化。除pH 3.0和3.5的处理组外,三者之间呈现一定的正相关性。     

悉生豚鼠体内复方双歧杆菌制剂对福氏志贺氏菌的作用

复方双歧杆菌制剂中,双歧杆菌(Bif.bifidum),乳酸杆菌(L.acidophilus),粪链球菌(Str.faecalis)三种菌都是人和动物肠道正常菌群之一,要研究它们和肠道致病菌之间的关系,在普通动物体内是无法观察到.在国内我们首次应用悉生豚鼠进行复方双歧杆菌制剂对福氏志贺氏菌作用的研究,发现在悉生豚鼠体内复方双歧杆菌制剂对感染后的豚鼠有较明显的保护作用,保护率为75%,实验组未获得保护的豚鼠死亡时间较对照组延长6～10天,而对照组豚鼠在感染后24～48小时全部死亡,解剖后在肠道分泌物中分离出福氏志贺氏菌.本实验证明:复方双歧杆菌制剂是治疗腹泻较好的生物制剂,尤其对因菌群失调而引起的腹泻更是理想的治疗方法。     

Sequence analysis of 16S rRNA genes amplified from two ribosomal RNA gene clusters of Bifidobacterium bifidum

Two rRNA gene clusters were detected in the genome of Bifidobacterium bifidum KCTC 3202^T using Southern blot analysis. To analyse the sequences of the 16S rRNA genes from rrnA and rrnB, 16S rDNAs were amplified by PCR using DNA fragments purified from gel slices containing each of the rRNA gene clusters. The amplified 16S rDNAs from rrnA and rrnB were cloned into vectors and three clones of each gene sequenced. The resultant sequences were confirmed by direct sequencing of the 16S rDNAs from rrnA and rrnB. Sequence differences were not found between rrnA and rrnB in 1488 bp of the 16S rRNA genes.     

双歧杆菌在家兔实验性肝坏死模型中的靶向增殖研究

目的 探讨双歧杆菌在海扶超声聚焦刀（HIFU）制作的肝凝固性坏死模型中的靶向性增殖的可能性。方法 用HIFU制作成家兔肝凝固性坏死模型,经耳缘静脉注射两歧双歧杆菌活菌,通过检测注射前后家兔体温、红细胞、白细胞和血小板等生理指标的变化和注射后主要器官的组织切片来评价双歧杆菌作为基因治疗载体系统的安全性,同时对肝坏死模型等组织切片还做了革兰染色检测,证明了双歧杆菌在坏死组织中的增殖。结果 双歧杆菌能够在HIFU制作的肝凝固性坏死模型中增殖、而这一增殖过程并不影响家兔的生命安全。结论 双歧杆菌作为肿瘤基因治疗的靶向载体系统是安全和可行的,HIFU制作的肝坏死模型可以用于双歧杆菌基因治疗的靶向载体系统的辅助研究。     

Longitudinal study of effects of oral dosage of Bifidobacterium bifidum G9‐1 on Japanese cedar pollen‐induced allergic nasal symptoms in guinea pigs

Previous studies using experimental animal models have reported the beneficial effects of probiotics on allergic responses; however, their long‐term effects on allergic nasal symptoms in clinical settings have not yet been elucidated in detail. In the present study, a guinea pig allergic rhinitis model involving repeated inhalation challenges with a natural allergen, Japanese cedar pollen, was used to examine the longitudinal effects of Bifidobacterium bifidum G9‐1 (BBG9‐1) on allergic nasal symptoms. BBG9‐1 was administered orally once a day. Amelioration of nasal blockage was consistently observed throughout the experimental period in the BBG9‐1‐treated group. Although challenge‐induced sneezing was not significantly inhibited in the BBG9‐1‐treated group, prolonged treatment with BBG9‐1 slightly reduced the frequency of sneezing. Antigen‐specific IgE antibody production was also not inhibited in the BBG9‐1‐treated group. Increases in the numbers of eosinophils and neutrophils in nasal cavity lavage fluid collected after pollen challenge were almost completely suppressed by BBG9‐1 treatment, whereas those in mast cell mediators, histamine and cysteinyl leukotrienes were not. In contrast, increases in the levels of nitric oxide metabolites were potently suppressed. Furthermore, prolonged BBG9‐1 treatment markedly suppressed exogenous leukotriene D₄‐induced nasal blockage. Thus, prolonged oral administration of BBG9‐1 suppresses Japanese cedar pollen‐induced allergic nasal symptoms. The inhibitory mechanisms responsible may involve reductions in the responsiveness of target organs, such as endothelial cells in nasal mucosal blood vessels, to chemical mediators.     

Bifunctional properties and characterization of a novel sialidase with esterase activity from Bifidobacterium bifidum

ABSTRACT

Sialidases catalyze the removal of terminal sialic acid from various complex carbohydrates. In the gastrointestinal tract, sialic acid is commonly found in the sugar chain of mucin, and many enteric commensals use mucin as a nutrient source. We previously identified two different sialidase genes in Bifidobacterium bifidum, and one was cloned and expressed as an extracellular protein designated as exo-α-sialidase SiaBb2. The other exo-α-sialidase gene (siabb1) from the same bifidobacterium encodes an extracellular protein (SiaBb1) consisting of 1795 amino acids with a molecular mass of 189 kDa. SiaBb1 possesses a catalytic domain that classifies this enzyme as a glycoside hydrolase family 33 member. SiaBb1 preferentially hydrolyzes α2,3-linked sialic acid over α2,6-linked sialic acid from sialoglycan, which is the same as SiaBb2. However, SiaBb1 has an SGNH hydrolase domain with sialate-O-acetylesterase activity and an N-terminal signal sequence and C-terminal transmembrane region. SiaBb1 is the first bifunctional sialidase identified with esterase activity.

Abbreviations: GalNAc: N-acetyl-D-galactosamine; Fuc: L-fucose; Gal: D-galactose